M13噬菌体多克隆抗体的制备与鉴定

目的：制备M13K07辅助噬菌体多克隆抗体。方法：将M13K07辅助噬菌体进行大量扩增，以此作为免疫原于1、3、5周进行兔免疫，收集0周和免疫后2、4、6、8周免疫血清，用间接ELISA法对免免疫血清的效价进行检测，进一步用免疫斑点实验检测免多克隆抗体与M13K07辅助噬菌体的特异性结合能力。结果：M13K07辅助噬菌体免疫兔可产生高效价的多克隆抗体，效价达1：3200，并能特异性识别M13K07辅助噬菌体。结论：成功制备了高效价的M13K07辅助噬菌体多克隆抗体，为噬菌体展示技术研究提供了检测抗体。

Preparation and identification of polyclonal antibodies against M13 phage

Objective： To prepare and characterize rabbit-anti M13K07 helper phage polyclonal antibodies. Methods： The helper phage was amplified and used to immunize rabbits to obtain polyclonal antibodies. Begin from the first week, the same immunization schedule of subcutaneous multipoint injection was repeated three times at 2 weeks intervals. Begin from 0 week, take blood serum collection from ear vein every other week. The immunogenicity of the M13K07 helper phage was detected with Dot Immunobinding Assay and indirect ELISA was used to demonstrate the titer of antiserum in the immunized rabbit. Results： It showed that the specific rabbit antibody against M13K07 helper phage could combined with M13K07 with Dot Immunobinding Assay. Indirect ELISA analysis showed that the helper page can stimulate high levels specific IgG antibody. Along with the increase of immunization times, antibody levels keep rising, reached the peak at the 6th week. The titer of the polyclonal antibody was approximately1：3 200. Conclusion： In this study, polyclonal antibodies against this M13K07 helper phage is successfully generated and such helper phage polyclonal antibodies is important reagentfor functional analysis for phage.